• Journal of Plant Biotechnology
• /
• v.36 no.1
• /
• pp.7-12
• /
• 2009

This study was conducted to establish the optimal suspension culture system for both the propagation of embryogenic cells (ECs) and the induction of somatic embryos (SEs) of Kalopanax septemlobus. The proliferation rate of ECs was reduced as the inoculum density was increased; the highest rate was obtained when 0.1 g/100 ml of cells was initially inoculated. According to the analysis of cell growth pattern and cell growth cycle (G1, Sand G2/M), the cell growth started in 5 days culture initiation, grew rapidly until 15 days and then decreased gradually. Distinctive changes of the cell growth cycle by the culture periods was also observed; the growth cycle was doubled from initial 5.6% to 11.7% of S stage in 5 days culture and then reached in stable stages again. Therefore, the results indicated that a 15-day-cycle was the optimal culture period for the propagation of the ECs through the suspension culture. Furthermore, the cell inoculum density was also important for the induction of SE; more than 65% of SEs at the torpedo stage was induced by using the low level of cell inoculum (0.5 g/L), while the higher inoculum densities were rapidly reduced the proportion of SEs at that stage. Although the higher inoculum density delayed the development of SE, it did not affect the proportion of SEs at the globular and heart stage. In conclusion, this study showed that the suspension culture of the Kalopanax septemlobus ECs through the control of inoculum density was an efficient way for both the propagation of ECs and the induction of SEs, suggesting that the development of this system might help to reduce the culture period for the somatic embryo production.

• Proceedings of the Korean Society of Grassland Science Conference
• /
• 1999.06a
• /
• pp.73.2-74
• /
• 1999

Orchardgrass의 종자배양 유래의 캘러스를 현탁배양하여 현탁배양기간별 부정배형성정도와 식물체 재분화율 등에 대한 몇 가지 실험을 수행한 바, 2주 간격으로 4회계 대배양 하였을 때 계대배양 횟수가 증가됨에 따라 식물체 재분화율이 증가되었다. 종자배양에서 형성된 캘러스의 현탁배양에서 모양이 둥근세포와 그들의 세포괴는 배양 30일 후에 최대치를 나타내었고, 그 이후는 감소하였다.(중략)

• Proceedings of the Korean Aquaculture Society Conference
• /
• 2003.10a
• /
• pp.88-89
• /
• 2003

Rotifer의 먹이 생물로 이용되는 Chlorella의 대량 배양에는 농업용 비료가 널리 이용되고 있다. 그러나 농업용 비료만을 사용할 경우 f/2배지에 비해 성장이 낮은 단점이 있다. 따라서 본 연구는 농업용 비료 배지에 여러 가지 미량 성분을 첨가하여 그 성장 변화를 측정하고, 고밀도 대량 배양에 필요한 미량 원소 성분을 파악하고자 하였다. 먼저 Chlorella의 대량 배양에 이용되는 농업용 비료 배지(복합, 요소비료)에 실내 배양에 주로 이용되는 f/2배지의 trace element 성분 중 코발트(Co), 구리(Cu), 아연(Zn) 및 몰리브덴(Mo)를 f/2배지의 함량을 기준으로 각각 0.5, 1.0, 1.5 및 2.0배로 첨가하여 $25^{\circ}C$, 5,000 lux 연속조명, 15$\textperthousand$에서 배양한 결과 각 미량 원소의 첨가 함량이 높을수록 Chlorella의 성장은 높게 나타났다. 전체 실험구 중에서 아연 2.0배, 구리 2.0배를 첨가한 비료 배지에서 S.G.R.이 각각 0.3598, 0.3599의 성장을 나타내었으며, 아연과 구리 성분을 첨가한 Chlorella에서 높은 성장률을 보였다. 위의 실험에서 첨가효과가 좋은 Cu와 Zn 2.0배와 철(Fe)과 망간(Mn)을 같은 비율(0.5~2.0배)로, 다시 배양한 결과, Cu 2.0배와 Zn 2.0배는 S.G.R.이 각각 0.6283과 0.6231로 가장 높았으며, Fe 2.0배, Mn 2.0배와 1.5배에서 S.G.R이 각각 0.6210, 0.6200, 0.6183의 높은 성장률을 나타내었다. 위의 실험에서 농업용 비료에 첨가된 모든 trace element 중에서 가장 좋은 성장을 나타낸 구리(Cu) 성분의 정확한 공급 농도를 파악하기 위하여 농업용 비료에 2, 3, 4 및 5배를 첨가하여 $25^{\circ}C$, 5,000 lux, 15$\textperthousand$에서 배양한 결과, S.G.R.이 CU 함량을 3배로 첨가하였을 경우 2716$\times$$10^4$cells/$m\ell$로 가장 높은 세포 밀도를 나타내었다. 또한 5종의 trace element (Fe, Mn, Zn, Cu, Co)를 혼합하여 비료 배지(1.25배)에 첨가하여 미량 원소간의 상호 작용에 대해서 알아 본 결과, Fe와 Mn 두가지 성분만을 농업용 비료 배지에 혼합하였을 경우 세포수가 2720$\times$$10^4$cells/$m\ell$로서 높은 성장을 나타내었다. 다음으로 Fe, Mn, Zn 및 Cu 4종을 모두 혼합하여 첨가한 실험구가 2608$\times$$10^4$ cells/$m\ell$ 높은 세포 밀도를 나타내었다. 이러한 결과로서 옥외에서 농업용 비료배지에 구리(Cu, f/2배지 기준 3.0배)성분을 첨가함으로써 Chlorella의 성장을 효율적으로 향상시킬 수 있을 것으로 판단된다.

• Korean Journal of Medicinal Crop Science
• /
• v.3 no.2
• /
• pp.100-106
• /
• 1995

This study was conducted to find the factors affecting somatic embryogenesis in suspension culture of Rehmania glutinosa and investigate the possibility of artificial seed production by encapsulation of somatic embryos. Linsmeier-Skoog medium was appeared as proper for somatic embryogenesis. Sucrose with $3{\sim}5%$ as carbon sources was good for somatic embryogenesis, and both ammonium and nitrate nitrogen were necesary for normal somatic embryo production. BA with NAA or kinetin with NAA were better than the use of cytokinin alone for both somatic embryogenesis and numbers of somatic embryos. $AgNO_3$ as protectant for vitrification of seedlings in vitro culture had no harmful effect on somatic embryos. Sphericity of encapsulated seeds was good at 3% gel of sodium alginate but germination was better at 2.5% sodium alginate level. Artificial seeds were germinated and developed normal shoots and roots under in vitro condition.

• Proceedings of the KSAR Conference
• /
• 2004.06a
• /
• pp.246-246
• /
• 2004

암모니아는 murine과 sheep의 난자의 체외 배양 시 배 발달과 착상, 태아 발달에 영향을 미친다고 보고되어져 있다. 본 연구는 한우 난포의 크기에 따른 암모니아 농도 측정과 체외 성숙 시간에 따라 발생되는 암모니아의 농도가 배 발달율에 미치는 영향을 검토하였다. 도축장 유래 한우 난소의 직경(3 ㎜∼30 ㎜)난포에서 난포액을 채취하였으며, 그리고 각각의 체외 성숙 시간에 따라 배양액을 회수하였다. 암모니아 농도 측정은 ammonia Kit를 이용 spectrophotometer로 630 ㎚에 측정하였다. (중략)

• Journal of Plant Biotechnology
• /
• v.37 no.4
• /
• pp.494-504
• /
• 2010

The effect of the addition of the fresh medium, volume of under solid medium in double layer culture as well as the medium change after pretreating microspores on the production of embryos in microspore culture of hot pepper (Capsicum annuum L.) has been studied. When cultured after heat pre-treatment, changing pretreatment media with fresh culture media proved to be more effective for embryo production rather than supplementing additional culture media. Heat-pretreating for 3 days turned out more effective for embryo production than pretreating for 1 or 2 days. In the case of anther pretreatment, the addition of fresh medium after culture was not effective for embryo production. In pretreating microspores, however, supplementing additional fresh culture media greatly improved embryo yield and quality. The best time point of media addition was 4 days after culture commenced, and the most effective number of times of media addition was one time addition. Moreover, the effective volume of added medium in double layer culture for embryo production was 1.5 ml. The addition of media more than 1.5 ml reduced both embryo yield and quality. Double layer medium was more effective for embryo development than liquid medium. When the volume of under solid medium increased ranging from 3 ml to 7 ml, more cotyledonary embryos were produced in either 5 ml or 7 ml compared to 3 ml, even though the total number of embryos were highest in 3 ml. These results can be used as an important data for establishing an efficient microspore culture system for producing high frequency of normal embryos in hot pepper.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.2
• /
• pp.85-90
• /
• 1994

Attempts were made to regenerate plants from petiole explane of Forest Pimpinella barchycarpa via repetitive somatic embryogenesis. Effective induction of somatic emb교ogenesis was achieved on both MS and modified $B_{5}\;(mB_{5})$ media containing BA + 2,4-D or BA + 2,4-D + NAA under light condition (16-h photoperiod/day) cultures. The explants exposed to the ligt produced numerous somatic embryos while those kept under the dark did not form any on the same medium. Somatic embryos at different developmental stages were observed to arise within a individual explants. Plantlets could be regenerated on $mB_{5}$ basal medium or $mB_{5}$ containing 0.1 mg/L NAA Secondary adventive embryos were formed on the surface of the somatic embryos. Therefore, repetitive somatic embryogenesis could be achieved by secondary embryogenesis. Although the treatment of 2,4-D or NAA alone was effective in callus formation and growth, but not in induction of somatic embryogenesis. Some explants, cultured on NAA-containing media in darkness, produced only adventive roots. The embryogenic potential was maintained for two years when subcultured to BA and 2,4-D containing media with 5 weeks inteval. Regenerated plantlets were maintained on $mB_{5}$ or MS basal media for 4 to 6 more weeks and transferred to soil of an artificial mixture for acclimation. Most plantlets (more than 97%) survived, and grew without any deformity.

• Journal of Plant Biology
• /
• v.28 no.2
• /
• pp.141-148
• /
• 1985

The effects of sizes and densities of cells cultured, conditioned medium, and media pH on the somatic embryogenesis of carrot (Daucus carota L.) were examined. A large number of globular embryoids was formed after 4 days in cell culture, and later globular embryoids developed into heart and torpedo shape. High cell density resulted in higher number and better growth of embryos, especially on conditioned medium than Murashige-Skoog medium. The fresh weight and number of embryoids formed increased with the decrease in cell size. The significant reduction in fresh weight and number of embryoids was obtained when culturing cells with diameter of over 90 ${\mu}{\textrm}{m}$. Dry weight and number of embryoids were markedly reduced with medium pH of 4 or 7, but promoted with pH 6.0.

• Horticultural Science & Technology
• /
• v.19 no.3
• /
• pp.378-383
• /
• 2001

Embryo culture, ovule culture and ovary slice culture were tested to find optimum method for overcoming post fertilization barrier in interspecific crosses between L. longiflorum 'Gelria' and L. cernuum. Although reciprocal crosses between the species were carried out by cut-style pollination method, fruits developed only in crosses of L. longiflorum${\times}$L. cernuum. On the 40 days after pollination, ovaries were sliced into 2-4mm thickness and cultured on a hormone-free Murashige-Skoog (MS) medium, supplemented with 2%, 4%, 6%, 8% and 10% sucrose. For the L. longiflorum Gelria'${\times}$L. cernuum cross, ovule development was found to be best at 6% sucrose and a lot of hybrid plant lets established directly from the ovary slice culture and subsequent ovule culture. High concentration of sucrose above 8% made ovules abort or vitrificate from 40 days after culture. In contrast, ovules from the L. cernuum${\times}$L. longiflorum 'Gelria' cross swelled well in ovary slice culture, however, they did not germinated in subsequent ovule culture. On the 60 days after pollination, ovules thicker than 0.6mm was interpreted as one containing embryo. The embryo size ranged from 1.2 mm to 1.7 mm, and in vitro development of the excised embryos was found to be best with the MS medium (pH 5.8), supplemented with $0.1-1 mg{\cdot}L^{-1}$ NAA and 6% sucrose. Thick ovules excised 60 days after pollination germinated about 60% as normal seeds in MS medium supplemented with 6% sucrose and free hormone. The ovule culture 60 days after pollination was concluded to be most recommendable to produce interspecific hybrids in large scale crosses between L. longiflorum 'Gelria' and L. cernuum by the reason of easy procedure.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.1
• /
• pp.51-56
• /
• 1998

This study was carried out in order to establish plant regeneration via somatic embryogenesis from leaf explant of Ostericum koreanum Kitagawa and to elucidate the effects of NAA and cytokinins (kinetin, BA) on the abnormalities of somatic embryo and the relationship between thecotyledon numberand germinability. Calli were formed on leaf explants cultured on MS agar medium supplemented with various concentrations (0, 0.1, 0.5, 1, 2 mg/L) of NAA and cytokinins. The calli were white, watery and soft, became browning during cultures. Somatic embryos were formed from pale yellowish calli derived browning calli. High frequency somatic embryos were observed on MS medium containing 1 mg/L NAA and 0.1 mg/L BA after 60 days of culture. The mature somatic embryos germinated into plantlets without subculture after 2 weeks. The frequency of normal somatic embryo with two cotyledons was 39.8%. On the other hand, cotyledonary abnormalities of somatic embryos were observed at considerable frequency: 33.6% of somatic embryo with one cotyledon, 15.3% cotyledons with three, 8.2% four cotyledons and 3.1% jar shaped cotyledon. Germination frequency of somatic embryos with two cotyledons was 97.4%, and that of the embryos with abnormal cotyledon was almost similar to that of embryos with two cotyledons, except jar shaped somatic embryos (33.3%).